Date of Award


Degree Name

Biological Sciences


College of Science

Type of Degree


Document Type


First Advisor

Charles C. Somerville

Second Advisor

Thomas K. Pauley

Third Advisor

Dan K. Evans


Aneides aeneus, the Green Salamander, is a plethodontid species ranging from southern Pennsylvania to central Alabama and from eastern Mississippi to Maryland. Populations of Green Salamanders most often inhabit cliffs and rocky outcrops which are moist but not wet and shaded. The animal is listed as a species of special concern by the West Virginia Division of Natural Resources Wildlife Diversity Program. Genetic and reproductive isolation due to strict habitat preferences of the Green Salamander could be a factor in the decline of the species. Seventeen tissue samples were retrieved from five sites within the Westvāco Wildlife and Ecosystem Research Forest (WWERF) in Randolph County, West Virginia for the purposes of determining the extent of gene flow between those 5 separate populations. The polymerase chain reaction was used to amplify approximately 940 base pairs of the ND4 NADH dehydrogenase region of mitochondrial DNA (mtDNA). These amplicons were sequenced and then analyzed using distance matrix phylograms, maximum parsimony cladograms, and linear regression analysis. Tree topologies, rooted by an outgroup Green Salamander specimen from North Carolina, showed no consistent grouping by individual populations. The North Carolina specimen consistently branched away from the West Virginia specimens. However, linear regression analysis showed a significant correlation (R=0.645, P=0.008) between genetic distances and geographical distances of the samples both within the WWERF and extending to the North Carolina specimen. This correlation was not significant for populations which were within one kilometer of each other. Linear regressions of protein genetic distances and geographic distances proved to have no significant correlation for populations inside the WWERF. We were unable to demonstrate, through phylogenetic methods or linear regressions, that populations within one kilometer are genetically isolated.This work suggests that gene flow can occur within a one kilometer radius, but is reduced or absent at three kilometers for A. aeneus.Continued monitoring of these populations and examination of the ND4 region of mtDNA from other populations is important for the Green Salamander. Knowledge of the extent of species migration is important to provide proper protective measures for this species.


Aneides - Migration - West Virginia.

Salamanders - West Virginia.

Westvaco Wildlife and Ecological Research Forest (Randolph Co., W. Va.)